Using tweezers, removed gill tissue from centrifuge tube and placed in weigh boat

Cut gill tissue in half with razor blade

Disposed of razor blade

Placed one gill tissue half back in original tube, the other in the tube labelled “1/2”

Put both tubes back in dry ice to stay cold

Tweezers cleaned by dipping in 10% bleach solution, and then rinsing in nanopure water

Repeated for all samples

When cutting the samples, I realized we forgot the step where we rinsed tweezers with nanopure AFTER we cleaned them with bleach! :open_mouth: I called Emma, and she said to dispose of those samples since they were possibly contaminated by bleach. Additionally, she said we should replace the bleach with ethanol, since trace elements of bleach in our samples would be problematic. Since I was dealing with all of my bare condition samples first, I replaced them with different samples.

Table 2. Revised samples for protein extraction. I wanted to use O127 for extractions but again, I couldn’t find it in the sample box. I replaced with O124.

Site

Condition

1

2

3

PG

B

O26

O60

O90

PG

E

O31

O71

N/A

FB

B

O66

O39

N/A

FB

E

O64

O46

O32

WB

B

O128

O124

O137

WB

E

O140

O145

O147

SK

B

O118

O100

N/A

SK

E

O103

O101

N/A

CI

B

O12

O14

O17

CI

E

O01

O08

N/A

The revised protocol:

Using tweezers, removed gill tissue from centrifuge tube and placed in weigh boat

Cut gill tissue in half with razor blade

Disposed of razor blade

Placed one gill tissue half back in original tube, the other in the tube labelled “1/2”

Put both tubes back in dry ice to stay cold

Tweezers cleaned by dipping in ethanol, and then rinsing in nanopure water

Repeated for all samples

I noticed that the amount of gill tissue in the original DNR sample vials varied between samples. Some had large chunks of tissue that were easily split, some barely had any tissue. I ran into something similar last time, so I don’t think it’s going to be much of an issue.

Made 50 mM NH4HCO3 + 6M urea solution

Protocol used can be found here, or viewed below. This solution must be used no later than 24 hours after it is made, or it is no longer viable.

Measured 10 mL of nanopure water in a graduated cylinder, and poured into falcon tube